Next Article in Journal
Rumen Fermentation and Fatty Acid Composition of Milk of Mid Lactating Dairy Cows Grazing Chicory and Ryegrass
Next Article in Special Issue
Gene Expression Profiling in Ovaries and Association Analyses Reveal HEP21 as a Candidate Gene for Sexual Maturity in Chickens
Previous Article in Journal
Evaluating the Reliability of Non-Specialist Observers in the Behavioural Assessment of Semi-Captive Asian Elephant Welfare
 
 
Search for Articles:
Title / Keyword
Author / Affiliation / Email
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
 
Journals
Animals
Volume 10
Issue 1
10.3390/ani10010168
animals-logo
Submit to this Journal Review for this Journal Propose a Special Issue
► Article Menu

Article Menu

share Share announcement Help format_quote Cite question_answer Discuss in SciProfiles thumb_up
...
Endorse textsms
...
Comment
first_page
settings
Order Article Reprints
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
Article

Myostatin (MSTN) Gene Indel Variation and Its Associations with Body Traits in Shaanbei White Cashmere Goat

by
Yi Bi
1,
Bo Feng
1,
Zhen Wang
1,2,3,
Haijing Zhu
2,3,
Lei Qu
2,3,
Xianyong Lan
1,
Chuanying Pan
1,* and
Xiaoyue Song
2,3,*
1
Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University, Yangling 712100, China
2
Shaanxi Provincial Engineering and Technology Research Center of Cashmere Goats, Yulin University, Yulin 719000, China
3
Life Science Research Center, Yulin University, Yulin 719000, China
*
Authors to whom correspondence should be addressed.
Animals 2020, 10(1), 168; https://doi.org/10.3390/ani10010168
Submission received: 26 November 2019 / Revised: 12 January 2020 / Accepted: 13 January 2020 / Published: 19 January 2020
(This article belongs to the Collection Genomics, Transcriptomics, and Computational Biology for Biodiversity Studies and Quality-Related Traits Selection in Livestock)
Browse Figures
Review Reports Versions Notes

Abstract

:

Simple Summary

As a major gene to regulate the muscle mass of animals, Myostatin (MSTN) plays a negative role in regulating the number and size of skeletal myocytes. Additionally, it is reported that mutations of MSTN gene contribute to the double-muscling (DBM) phenomenon. Therefore, mutations of MSTN gene will always be one of the hot spots. A 5 bp indel (c. –120 ins) in the 5’ untranslated region (5’ UTR) of goat MSTN gene was reported to relate to the growth traits of goat. However, all the sample sizes were limited. Herein, this study firstly enlarged the sample size (n = 1074, Shaanbei White Cashmere goat) to uncover the indel location, as well as its association with growing performance. Based on the data, the 5 bp insertion mutation in goat MSTN gene was significantly associated with the body height, height at hip cross, and chest width index in SBWC (p < 0.05), hinted that this insertion could be assigned to an effective molecular marker for growth trait in goat rearing.

Abstract

Myostatin (MSTN) gene, also known as growth differentiation factor 8 (GDF8), is a member of the transforming growth factor-beta super-family and plays a negative role in muscle development. It acts as key points during pre- and post-natal life of amniotes that ultimately determine the overall muscle mass of animals. There are several studies that concentrate on the effect of a 5 bp insertion/deletion (indel) within the 5’ untranslated region (5’ UTR) of goat MSTN gene in goats. However, almost all sample sizes were below 150 individuals. Only in Boer goats, the sample sizes reached 482. Hence, whether the 5 bp indel was still associated with the growth traits of goats in large sample sizes which were more reliable is not clear. To find an effective and dependable DNA marker for goat rearing, we first enlarged the sample sizes (n = 1074, Shaanbei White Cashmere goat) which would enhance the robustness of the analysis and did the association analyses between the 5 bp indel and growth traits. Results uncovered that the 5 bp indel was significantly related to body height, height at hip cross, and chest width index (p < 0.05). In addition, individuals with DD genotype had a superior growing performance than those with the ID genotype. These findings suggested that the 5 bp indel in MSTN gene are significantly associated with growth traits and the specific genotype might be promising for maker-assisted selection (MAS) of goats.

1. Introduction

Increasing need for mutton requires that certain growth-related traits of local goat breeds which are always of primary concern should be remarkably improved [1]. Early growth traits of animals, being regulated by multi-genes, as well as their interactions, play a vital role in influencing profitability in any meat-producing enterprise [2,3]. Thus, to establish optimal breeding programs, plenty of candidate genes, as well as their genetic variations should be appropriately used [4,5,6,7]. Maker-assisted selection (MAS) can satisfy the need to screen crucial genes and consider the relation between their polymorphisms and growth-related traits [8,9,10,11]. Herein, establishing a MAS system and performing candidate genes analyses will speed up the development of goat breeding [12,13,14,15].
Myostatin (MSTN) gene, also known as growth and differentiation factor 8 (GDF8), is highly conserved in many tissues (including the mammary gland) but most prominently in skeletal muscle [16]. MSTN exons encode for a 375 amino-acid (aa) latent protein that undergoes significant post-translational modification to become biologically active [17]. It is one of the predominant regulators that inhibits skeletal muscle growth especially during the process of pre-natal muscle development which completely determines the number of myofibers: Muscle precursor proliferation, myoblast proliferation, and differentiation [18,19]. The MSTN gene upregulates the cyclin-dependent kinase inhibitor p21 expression (Waf1, Cip1 signal pathway) and downregulates the Cdk2 protein which contributes to the arrest of myoblast in G (1)-phase of cell cycle. Ultimately, the deregulated of myoblast proliferation leads to muscular hyperplasia [20]. The deficiency of MSTN contributes to an increase in skeletal muscle mass known as double-muscling (DBM). The double-muscle phenomenon was firstly reported by Culley and Woodfall in 1807 [21]. The extensive breakthrough in understanding skeletal myogenesis was accomplished by delineating natural mutation within the MSTN causing double muscling phenotype in the Belgian Blue and Piedmontese Cattle breeds [22,23,24,25]. A similar phenotype can also be observed in the MSTN gene of knockdown animals. The first MSTN-deficient mice were reported to be 30% greater in muscle mass than heterozygous and wild-type normal mice [26,27]. With the lucubration of the above phenomenon, RNAi, molecular cloning, gene knockdown, as well as gene editing technology such as CRISPR/Cas9, TALEN, and ZFN were also applied to the goat sheep [28,29,30,31,32,33,34].
In addition, the MSTN mutations are necessary for hypertrophy, muscle mass, growth, and other traits, with lesser roles played by other gene variants [35]. Therefore, MSTN and its variation are becoming one of the hot spots for association analysis of growth traits and meat traits in the animal breeding. Numerous studies and researches reported that there were various SNPs and indels loci in both domestic and exotic commercial breeds of goat, sheep, cattle, pig, rabbit, as well as poultry [36]. Additionally, plenty of research focused on a 5 bp indel in 5’ UTR of MSTN gene within different goat breeds to uncover its distributions and effects on growth traits of goats [37]. A research detected distributions of the 5 bp indel in Black Bengal, Sirohi, Osmanabadi, Jakhrana, Jamunapari, Barbari, Marwari goat breeds [38]. Furthermore, several reports revealed that the 5 bp indel was associated with body length in Boer goats [39] and circumference of cannon bone, hip width, chest depth in backcrossed offsprings of Boer goats and Tangshan Dairy goats [40]. What is worth mentioning here are the relevant reports about the association between variations and growth traits that are limited in sample sizes [39,40].
As the main genetic source of cashmere in local goat industry of North China, the Shaanbei White Cashmere goat which is a hybrid goat between Liaoning Cashmere goat (♂) and Shaanbei Ziwuling Black goat (♀), has the resistance to the crude feed, cold, wind-blown sand and disease [41]. Additionally, it has superior qualities and quantities of cashmere than other cashmere goat breeds. However, growth traits of Shaanbei White Cashmere goats need improving. Rapid, accurate, as well as an effective method of MAS is presented in [42,43,44]. Herein, this study aimed at finding an effective and faithful DNA maker which could accelerate the process of selective breeding in goat rearing. A total of 1074 Shaanbei White Cashmere goats were analyzed, aiming at enhancing the robustness, as well as revealing the impact of the 5 bp indel on the growth performance. These results may provide references for further research on applying MAS to the goat industry.

2. Materials and Methods

All experiments implemented in this study were approved by the International Animal Care and Use Committee of the Northwest A&F University (IACUC-NWAFU) and fully followed local animal welfare guidelines, laws, and policies. Additionally, all experimental processing in this study were approved by the International Animal Care and Use Committee (IACUC) of the Northwest A&F University (protocol number NWAFAC1008) on animal experiment, including sample collection, were performed with the guidelines of the ethics commission.

2.1. Sample Collection, Phenotypic Traits Recording, and Genomic DNA Isolation

A total of 1074 ear tissue samples (adults n = 472; kids, n = 602) were collected randomly from the Shaanbei White Cashmere goat breeding farm. Nine growth trait records (some body weight records were missing) were provided by the agricultural technical station of Yulin city, Shaanxi Province. DNA was extracted using a high-salt extraction method [45,46]. After being assayed by a Nano-drop 1000 spectrophotometer (Thermo Scientific, Waltham, MA, USA), all DNA samples were diluted to the same standard of 20 ng/μL and were stored at 4 °C temporarily. Additionally, 30 DNA samples of Shaanbei White Cashmere goat were mixed in a genomic DNA pool for a polymerase chain reaction (PCR) analysis [47].

2.2. Primer Design, PCR Amplification, and Genotyping

Using the Primer Premier 6.0 (Premier, Canada), the primer pair (F: 5’-ACTGGTGTGGCAAGTTGTCTCT-3’; R: 5’-TTCCTTCTGCTCGCTGTTCTCA-3’) for amplification of indel loci in 5’ UTR was designed based on the goat MSTN gene sequence (NC_030809.1) and the Ensemble Indel-database (http://apr2019.archive.ensembl.org/index.html/indel). Assays were performed by touch-down PCR in a 13 μL volume, containing 6.5 μL 2 × mix, 0.2 μL each of forward and reverse primers, 0.5 μL genomic DNA (20 ng/μL), and 5.6 μL ddH2O. The PCR protocol was as below: Initial denaturation for 5 min at 95 °C; followed by 18 cycles of denaturation for 30 s at 95 °C, annealing for 30 s at 68 °C (with a decrease of 1 °C per cycle), extension for 15 s at 72 °C; another 38 cycles of 30 s at 95 °C, 30 s at 50 °C, and 15 s at 72 °C, and a final extension for 5 min at 72 °C, with subsequent cooling to 12 °C. Using 3.5% agarose gel electrophoresis, spoting 5 μL and running for 1.5 h, the 5 bp indel of goat MSTN gene was examined via electrophoresis at 120 V voltage. Then, genotypes were identified when combined with sequencing.

2.3. Statistical Analyses

To explore the genetic diversity of the indel variants in the investigated goat population, we calculated genetic parameters. The genotype and allele frequencies reflect the genetic composition of the indel variant in the tested goat population. We calculate the population genetic diversity indices, including homozygosity (Homo), heterozygosity (Hetero), and polymorphism information content (PIC) in Nei’s methods [48]. Homo and Hetero (Homo + Hetero = 1) are measures of genetic variation of a population. PIC is an indicator of polymorphism. Based on PIC values, the genetic variations classified as low genetic diversity (PIC < 0.25), medium genetic diversity (0.25 < PIC < 0.5), and high genetic diversity (PIC > 0.5) [49]. Cause there were only two genotypes (DD and ID), the independent-samples t-test was used in the study [50].

3. Results

3.1. Identification and Genetic Parameter Analysis of Indel Variation

A 5 bp indel (c. −120 ins) in the 5’ UTR of goat MSTN gene was detected. There were two different genotypes: Homozygote deletion type (deletion/deletion: DD, 206 bp); heterozygote type (insertion/deletion: ID, 211 bp and 206 bp) (Figure 1).
The genotype and allele frequencies, as well as other genetic parameters, associated with the MSTN indel loci were calculated to determine the genotype distribution among Shaanbei White Cashmere goats (Table 1). The data indicated that “D” allele (0.943) of the 5 bp indel was more frequent than “I” allele (0.057). Based on PIC values, the 5 bp locus had a low genetic diversity (PIC = 0.055).
Published reports also performed the genetic parameter analyses in several different goat breeds (Table S1). The “D” alleles were totally more frequent than “I” alleles while different goat breeds had different genetic diversities.

3.2. Associations between Indel Variations and Growth Traits

From (Table 2 and Table S2, Figure 2), association analyses showed that the 5 bp indel was significantly associated with the body height, height at hip cross and chest width index (p < 0.05) from 602 kids. Interestingly, the “DD” genotype was superior than “ID” genotype in the body height and the height at hip cross which was reverse to the chest width index.
In addition, we did a correlation analysis between growth traits of which “DD” genotype had a superior performance than “ID” genotype (BH, HHC, CC, BW). Results manifested that analyzed growth traits were positively correlated with each other (Table S3).
There is no significance between the 5 bp indel and growth traits in adults. However, the individual with “DD” had a superior performance than those with “ID” in body height, height at hip cross, hip width, chest width, cannon circumference, chest depth, body weight, heart girth traits which was coincident with the body height and height at hip cross of the kids.

4. Discussion

In this study, the relation between the 5 bp indel and growth traits of SBWC goats was detected. Genetic parameter analysis confirmed that the 5 bp indel was at Hardy–Weinberg equilibrium. Additionally, the allelic frequency of “D” is higher than “I” which was consistent with several exotic goat breeds such as Black Bengal, Sirohi, Osmanabdi, Maerhoz, and domestic goat breeds such as Boer, Matou goats. Interestingly, the “DD” genotype was the dominant genotype and had a superior phenotype than the “ID” genotype in the above goat breeds [38]. While the circumstance in Nubi goat breeds was different. It is reported that three genotypes of the 5 bp indel were found in the Nubi goat and the “I” allele, as well as the “ID” genotype was dominant in Nubi goat breed [39]. Additionally, the 5 bp indel within the Shaanbei White Cashmere goat was at low genetic diversity, while there were low, medium, and high genetic diversity in other domestic and exotic commercial goat breeds, respectively. In detail, in Black Bengal, Osmanabadi, Jakhrana, Jamunapari, Barbari, Marwari, Markhoz, Matongoats goat breeds, the 5 bp indel had low genetic diversities (0 < PIC < 0.164). In Boer, Haimen, Nubi, South group, North group, Foreign group, as well as the backcrossed offspring, the PIC values ranged from 0.279 to 0.375 which illustrated that the 5 bp indel among these breeds had medium genetic diversities. Only in Sirohi goats, the 5 bp indel had high genetic diversity (PIC = 1.133) [38,39]. Next, the association between different genotypes and growth traits was analyzed. The circumstance was that the 5 bp indel was distinctly related with body height, height at hip cross, and chest depth index in kids. It could be explained that the MSTN gene mutations gave rise to hyperplasia and hypertrophy which leaded to an increasing in muscle mass [51]. Additionally, we did a correlation analysis between growth traits which had a consistent phenotype with the body height and height at hip cross. It revealed that all growth traits had a positive correlation (p < 0.05) with each other. Hence, we speculate that the 5 bp indel can affect other growth traits of Shaanbei White Cashmere goats.
The MSTN gene plays a crucial role in the development of domestic animals due to its key function in muscularity [52]. Its exons code for a 375 amino-acid (aa) latent protein which will become biologically active undergoing significant post-translational modification. Firstly, through formatting the disulphide bonds, the polypeptide undergoes intracellular homodimerization which cleaved the polypeptide to form the N-terminal propeptide region and the C-terminal mature region. The 12 kDa C-terminal mature fragment of MSTN gene initiates an intracellular signaling cascade according to bind and activates the activin type II receptor at the cell surface (ActRIIB and to a lesser extent ActRIIA). Subsequent autophosphorylation of ActRIIB gives rise to recruiting and activating the low affinity type I receptor for activin ALK-4 or ALK-5. Moreover, activated type I receptor kinase phosphorylates the transcription factors Smad2 and Smad3 which allows them to interact with Smad4 (co-Smad) and to translocate to the nucleus to activate target gene transcription. Importantly, the activation of the MSTN receptor also inhibits Akt (protein kinase B) activity which displays a decisive role in muscle protein synthesis and cell proliferation [53]. On the contrary, the deletion of goat MSTN gene will enhance the muscle protein synthesis and cell proliferation which contribute to an aggrandizement on the mass of muscle, then gain the body weight. Since the body height had a positive correlation with the body weight [54], the deletion of the MSTN gene was considered to have a positive effect on body weight. Additionally, we also hypothesized that the mutations of goat MSTN gene were related to the phenotype of other growth traits.
Briefly, the 5 bp indel of 5’ UTR within the goat MSTN was found to show different genotypic and allelic distributions in Shaanbei White Cashmere goats which manifest that it can be potential DNA makers for a further step of selecting high-quality individuals with MAS in breeding.

5. Conclusions

The 5 bp insertion mutation in the 5‘ UTR within the MSTN gene was detected in SBWC goats. Additionally, the “D” allele was the dominant allele in this population which displayed a low genetic diversity. Interestingly, the “DD” genotype was the dominant genotype. Furthermore, association analysis showed that the 5 bp indel was significantly associated with the body height, height at hip cross, and chest width index in SBWC (p < 0.05), hinting that this insertion could be assigned to an effective molecular marker for growth trait in goat rearing.

Supplementary Materials

The following are available online at https://www.mdpi.com/2076-2615/10/1/168/s1. Table S1. Genetic parameters of the 5 bp indel within GDF9 gene in different goat breeds; Table S2. Relationship between the 5 bp indel locus within the MSTN gene and the growth traits in SBWC adults (LSM a ± SE) (p < 0.05); Table S3. Correlation matrix of different growth traits in SBWC.

Author Contributions

Conceptualization, Y.B., B.F., X.S., C.P.; methodology, Y.B., Z.W.; software, Y.B.; validation, Z.W., X.L.; formal analysis, Y.B., B.F., Z.W.; investigation, X.L.; resources, H.Z., L.Q., X.L., X.S., C.P.; data curation, X.L.; writing—original draft preparation, Y.B.; writing—review and editing, Y.B., X.L., X.S., C.P.; visualization, C.P.; supervision, Y.B., X.L., X.S., C.P.; project administration, H.Z., L.Q., X.L., X.S., C.P.; funding acquisition, H.Z., L.Q., X.L., X.S., C.P. All authors have read and agreed to the published version of the manuscript.

Funding

This work was funded by the Shaanxi Provincial Scientific and Technological Innovation Project for Undergraduate of Northwest A&F University (S201910712052), the Scientific Research Foundation of the Education Department of Shaanxi Province (No. 19JK1004), the Young Talent Fund of University Association for Science and Technology in Shaanxi (No. 20190208), and the Research Foundation for High-Level Talents of Yulin University (No. 17GK22).

Acknowledgments

We are grateful to the staff at the Shaanbei White Cashmere (SBWC) goat breeding farm for allowing us to collect samples and for the support from the Agricultural Technical Station of Veterinary and Animal Husbandry of Yulin City, Shaanxi Province, P.R. China. We also greatly appreciated the Life Science Research Core Services (LSRCS) of Northwest A&F University (Northern Campus) for providing us with the platform.

Conflicts of Interest

We confirm that there is no conflict of interest with any financial organization for the material discussed in the manuscript, and the publication of the manuscript is approved by all authors.

References

  1. Zhang, Z.J.; Ling, Y.H.; Wang, L.J.; Hang, Y.F.; Guo, X.F.; Zhang, Y.H.; Ding, J.P.; Zhang, X.R. Polymorphisms of the myostatin gene (MSTN) and its relationship with growth traits in goat breeds. Genet. Mol. Res. 2013, 12, 965–971. [Google Scholar] [CrossRef] [PubMed]
  2. Dalvit, C.; De Marchi, M.; Cassandro, M. Genetic traceability of livestock products: A review. Meat Sci. 2007, 77, 437–449. [Google Scholar] [CrossRef] [PubMed]
  3. Warner, R.D.; Greenwood, P.L.; Pethick, D.W.; Ferguson, D.M. Genetic and environmental effects on meat quality. Meat Sci. 2010, 86, 171–183. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  4. Zhou, D.K.; Yang, B.G.; Duan, X.H.; Na, R.S.; Han, Y.G.; Zeng, Y. Association analysis of sixty-seven single nucleotide polymorphisms with litter size in Dazu black goats. Anim. Genet. 2019. [Google Scholar] [CrossRef]
  5. Gao, Y.; Huang, B.; Bai, F.; Wu, F.; Zhou, Z.; Lai, Z.; Li, S.; Qu, K.; Jia, Y.; Lei, C.; et al. Two novel SNPs in RET gene are associated with cattle body measurement traits. Animals 2019, 9, 836. [Google Scholar] [CrossRef] [Green Version]
  6. Shi, B.; Jiang, Y.; Chen, Y.; Zhao, Z.; Zhou, H.; Luo, Y.; Hu, J.; Hickford, J.G.H. Variation in the Fatty Acid Synthase Gene (FASN) and Its Association with milk traits in gannan yaks. Aniamls 2019, 9, 613. [Google Scholar] [CrossRef] [Green Version]
  7. Lee, J.; Kim, J.M.; Garrick, D.J. Increasing the accuracy of genomic prediction in pure-bred Limousin beef cattle by including cross-bred Limousin data and accounting for an F94L variant in MSTN. Anim. Genet. 2019. [Google Scholar] [CrossRef]
  8. E, G.X.; Duan, X.H.; Zhang, J.H.; Huang, Y.F.; Zhao, Y.J.; Na, R.S.; Zhao, Z.Q.; Ma, Y.H.; Chu, M.X.; Basang, W.D.; et al. Genome-wide selection signatures analysis of litter size in Dazu black goats using single-nucleotide polymorphism. 3 Biotech. 2019. [Google Scholar] [CrossRef]
  9. Kang, Z.; Zhang, S.; He, L.; Zhu, H.; Wang, Z.; Yan, H.; Huang, Y.; Dang, R.; Lei, C.; Chen, H.; et al. A 14-bp functional deletion within the CMTM2 gene is significantly associated with litter size in goat. Theriogenology 2019, 139, 49–57. [Google Scholar] [CrossRef]
  10. La, Y.; Liu, Q.; Zhang, L.; Chu, M. Single nucleotide polymorphisms in SLC5A1, CCNA1, and ABCC1 and the association with litter size in small-tail Han sheep. Animals 2019, 9, 432. [Google Scholar] [CrossRef] [Green Version]
  11. Zhou, M.; Pan, Z.; Cao, X.; Guo, X.; He, X.; Sun, Q.; Di, R.; Hu, W.; Wang, X.; Zhang, X.; et al. Single nucleotide polymorphisms in the HIRA gene affect litter size in small tail Han sheep. Animals 2018, 8, 71. [Google Scholar] [CrossRef] [Green Version]
  12. Goshu, H.A.; Wu, X.Y.; Chu, M.; Bao, P.J.; Ding, X.Z.; Yan, P. Copy number variations of KLF6 modulate gene transcription and growth traits in Chinese Datong yak (Bos Grunniens). Animals 2018, 8, 145. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  13. Li, W.Y.; Liu, D.L.; Tang, S.Q.; Tan, X.X.; Han, R.L.; Tian, Y.D.; Li, H.; Li, G.X.; Li, W.T.; Liu, X.J.; et al. A multiallelic indel in the promoter region of the Cyclin-dependent kinase inhibitor 3 gene is significantly associated with body weight and carcass traits in chickens. Poult. Sci. 2019, 98, 556–565. [Google Scholar] [CrossRef] [PubMed]
  14. Wang, J.Q.; Zhou, H.T.; Luo, Y.; Zhao, M.L.; Gong, H.; Hao, Z.Y.; Hu, J.; Hickford, J.G.H. Variation in the caprine KAP24-1 gene affects cashmere fibre diameter. Animals 2019, 9, 15. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  15. Ren, T.H.; Li, W.Y.; Liu, D.L.; Liang, K.; Wang, X.G.; Li, H.; Jiang, R.R.; Tian, Y.D.; Kang, X.T.; Li, Z.J. Two indels mutation in the promoter region of the QPCTL gene is significantly associated with body weight and carcass traits in chickens. Anim. Genet. 2019, 50, 279–282. [Google Scholar] [CrossRef]
  16. Ji, S.; Losinski, R.L.; Cornelius, S.G.; Frank, G.R.; Willis, G.M.; Gerrard, D.E.; Depreux, F.F.; Spurlock, M.E. Myostatin expression in porcine tissues: Tissue specificity and developmental and postnatal regulation. Am. Physiol. Soc. 1998, 275, 1265–1273. [Google Scholar] [CrossRef]
  17. Wolfman, N.M.; McPherron, A.C.; Pappano, W.N.; Davies, M.V.; Song, K.; Tomkinson, K.N.; Wright, J.F.; Zhao, L.; Sebaid, S.M.; Greenspan, D.S.; et al. Activation of latent myostatin by the BMP-1/tolloid family of metalloproteinases. Proc. Natl. Acad. Sci. USA 2003, 100, 15842–15846. [Google Scholar] [CrossRef] [Green Version]
  18. Beyer, T.A.; Narimatsu, M.; Weiss, A.; David, L.; Wrana, J.L. The TGFβ superfamily in stem cell biology and early mammalian embryonic development. Biochim. Biophys. Acta 2013, 1830, 2268–2279. [Google Scholar] [CrossRef]
  19. Liu, H.H.; Mao, H.G.; Dong, X.Y.; Cao, H.Y.; Liu, K.; Yin, Z.Z. Expression of MSTN gene and its correlation with pectoralis muscle fiber traits in the domestic pigeons (Columba livia). Poult. Sci. 2019. [Google Scholar] [CrossRef]
  20. Thomas, M.; Langley, B.; Berry, C.; Sharma, M.; Kirk, S.; Bass, J.; Kambadur, R. Myostatin, a negative regulator of muscle growth, functions by inhibiting myoblast proliferation. J. Biol. Chem. 2000, 275, 40235–40243. [Google Scholar] [CrossRef] [Green Version]
  21. Kambadur, R.; Sharma, M.; Smith, T.P.; Bass, J.J. Mutations in myostatin (GDF8) in Double-Muscled Belgian Blue and Piedmontese Cattle. Genome Res. 2015, 9, 910–916. [Google Scholar]
  22. Kambadur, R.; Sharma, M.; Smith, T.P.; Bass, J.J. Mutations in myostatin (GDF8) in double-musclód Belgian Blue and Piedmontese cattle. Genome Res. 1997, 7, 910–916. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  23. Grobet, L.; Martin, L.; Jcelet, D.; Pirottin, D.; Brouwers, B.; Riquet, J.; Schoeberlein, A.; Dunner, S.; Ménissier, F.; Massabanda, J.; et al. A deletion in the bovine myostatin gene causes the double-muscled phenotype in cattle. Nat. Genet. 1997, 17, 71–74. [Google Scholar] [CrossRef] [PubMed]
  24. Grobet, L.; Poncelet, D.; Royo, L.J.; Brouwers, B.; Pirottin, D.; Michaux, C.; Menissier, F.; Zanotti, M.; Dunner, S.; Georges, M. Molecular definition of an allelic series of mutations disrupting the myostatin function and causing double- muscling in cattle. Mamm. Genome 1998, 9, 210–213. [Google Scholar] [CrossRef] [PubMed]
  25. Marchitelli, C.; Savarese, M.C.; Crisa, A.; Nardone, A.; Marsan, P.A.; Valentini, A. Double muscling in Marchigiana beef breed is caused by a stop codon in the third exon of myostatin gene. Mamm. Genome 2003, 14, 392–395. [Google Scholar] [CrossRef]
  26. Grobet, L.; Pirottin, D.; Farnir, F.; Poncelet, D.; Royo, L.J.; Brouwers, B.; Christians, E.; Desmecht, D.; Coignoul, F.; Kahn, R.; et al. Modulating skeletal muscle mass by postnatal, muscle- specific inactivation of the myostatin gene. Genesis 2003, 35, 227–238. [Google Scholar] [CrossRef]
  27. Tang, L.; Yan, Z.; Wan, Y.; Han, W.; Zhang, Y. Myostatin DNA vaccine increases skeletal muscle mass and endurance in mice. Muscle Nerv. 2007, 36, 342–348. [Google Scholar] [CrossRef]
  28. Amrutlal, K.; Patel, A.K.; Tripathi, U.A.; Shah, R.K.; Joshi, C.G. Myostatin knockdown and its effect on myogenic gene expression program in stably transfected goat myoblasts. In Vitro Cell. Dev. Biol. Anim. 2014, 50, 587–596. [Google Scholar]
  29. Guo, X.; Wang, X.; Liang, B.; Di, R.; Liu, Q.; Hu, W.; He, X.; Zhang, J.; Zhang, X.; Chu, M. Differential expression of circular RNAs in polytocous and monotocous uters during the reproductive cycle of sheep. Animals 2018, 8, 160. [Google Scholar] [CrossRef] [Green Version]
  30. Hemlata, J.; Sanjeev, S.; Megha, K.; Sarkhel, B.C. Knockdown of the myostatin gene by RNA interference in caprine fibroblast cells. J. Biotechnol. 2010, 145, 99–102. [Google Scholar]
  31. Wang, X.L.; Yu, H.H.; Lei, A.M.; Zhou, J.K.; Zeng, W.X.; Zhu, H.J.; Dong, Z.M.; Niu, Y.Y.; Shi, B.B.; Cai, B.; et al. Generation of gene-modified goats targeting MSTN and FGF5 via zygote injection of CRISPR/Cas9 system. Sci. Rep. 2015, 5, 13878. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  32. Wang, X.; Niu, Y.; Zhou, J.; Zhu, J.; Ma, B.; Yu, H.; Yan, H.; Hua, J.; Huang, X.; Qu, L.; et al. CRISPR/Cas9-mediated MSTN disruption and heritable mutagenesis in goats causes increased body mass. Anim. Genet. 2017, 49, 43–51. [Google Scholar] [CrossRef] [PubMed]
  33. Yu, B.L.; Lu, R.; Yuan, Y.G.; Zhang, T.; Song, S.Z.; Qi, Z.Q.; Shao, B.; Zhu, M.M.; Mi, F.; Cheng, Y. Efficient TALEN-mediated myostatin gene editing in goats. BMC Dev. Biol. 2016, 16, 26. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  34. Lv, Q.; Yuan, L.; Deng, J.; Chen, M.; Wang, Y.; Zeng, L.Z.; Lai, L. Efficient generation of myostatin gene mutated rabbit by CRISPR/Cas9. Sci. Rep. 2016, 26, 25029. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  35. Kobolák, J.; Gócza, E. The role of the myostatin protein in meat quality: A review. Arch. Anim. Breed. 2002, 45, 159–170. [Google Scholar] [CrossRef]
  36. Aiello, D.; Patel, K.; Lasagna, G. The myostatin gene: An overview of mechanisms of action and its relevance to livestock animals. Anim. Genet. 2018, 49, 505–519. [Google Scholar] [CrossRef] [Green Version]
  37. Khani, K.; Abdolmohammadi, A.; Foroutanifar, S.; Zebarjadi, A. Assessment of polymorphisms in mysostatin gene and their allele substitution effects showed weak association with growth traits in Iranian Markhoz goats. J. Agric. Sci. 2017, 155, 519–526. [Google Scholar] [CrossRef]
  38. Singh, S.P.; Kumar, R.; Kumar, S.; Mitra, A. Characterization of 5’ upsteam region and investigation of TTTTA deletion in 5’UTR of myostatin (MSTN) gene in Indian goat breeds. Anim. Biotechnol. 2014, 25, 55–68. [Google Scholar] [CrossRef]
  39. Zhang, C.Y.; Liu, Y.; Xu, D.Q.; Wen, Q.Y.; Li, X.; Zhang, W.M.; Yang, L.G. Polymorphisms of myostatin gene (MSTN) in four goat breeds and their effects on Boer goat growth performance. Mol. Biol. Rep. 2012, 39, 3081–3087. [Google Scholar] [CrossRef]
  40. Li, X.L.; Liu, Z.Z.; Zhou, R.Y.; Zheng, G.R.; Gong, Y.F.; Li, L.H. Deletion of TTTTA in 5’UTR of goat MSTN gene and its distribution in different population groups and genetic effect on body weight at different ages. Front. Agric. China 2008, 2, 103–109. [Google Scholar] [CrossRef]
  41. Liu, A.Y. Progress in the production performance of Shaanbei white Cashmere goat. China Herbiv. Sci. 2018, 27, 449–455. [Google Scholar]
  42. Wu, X.; Zhou, X.; Ding, X.; Chu, M.; Liang, C.; Pei, J.; Xiong, L.; Bao, P.; Guo, X.; Yan, P. The selection of reference genes for quantitative real-time PCR in the ashidan yak mammary gland during lactation and dry period. Animals 2019, 9, 943. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  43. Yin, H.; Du, X.; Li, Q.; Pan, Z.; Wu, W.; Liu, H.; Li, Q. Variants in BMP7 and BMP15 3’-UTRs associated with reproductive traits in a large white pig population. Animals 2019, 9, 905. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  44. Yuan, Z.; Zhang, J.; Li, W.; Wang, W.; Li, F.; Yue, X. Association of Polymorphisms in candidate genes with the litter size in two sheep breeds. Animals. 2019, 9, 958. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  45. Aljanabi, S.M.; Martinez, I. Universal and rapid salt-extraction of high-quality genomic DNA for PCR-based techniques. Nucleic Acids Res. 1997, 25, 4692–4693. [Google Scholar] [CrossRef] [PubMed]
  46. Zhu, H.; Zhang, Y.; Bai, Y.; Yang, H.; Yan, H.; Liu, J.; Shi, L.; Song, X.; Li, L.; Dong, S.; et al. Relationship between SNPs of POU1F1 gene and litter size and growth traits in Shaanbei white cashmere goats. Animals 2019, 9, 114. [Google Scholar] [CrossRef] [Green Version]
  47. Chen, M.; Yan, H.; Wang, K.; Cui, Y.; Chen, R.; Liu, W.; Zhu, H.; Qu, L.; Pan, C. Goat SPEF2: Expression profile, indel variants identification and association analysis with litter size. Theriogenology 2019, 139, 147–155. [Google Scholar] [CrossRef]
  48. Nei, M.; Roychoudhury, A.K. Sampling variances of heterozygosity and genetic distance. Genetics 1974, 76, 379–390. [Google Scholar]
  49. Botstein, D.; White, R.L.; Skolnick, M.; Davis, R.W. Construction of a genetic linkage map in man using restriction fragment length polymorphisms. Am. J. Hum. Genet. 1980, 32, 314–331. [Google Scholar]
  50. Wang, Z.; Zhang, X.L.; Jiang, E.H.; Yan, H.L.; Zhu, H.J.; Chen, H.; Liu, J.W.; Qu, L.; Pan, C.Y.; Lan, X.Y. InDels within caprine IGF2BP1 intron 2 and the 3’-untranslated regions are associated with goat growth traits. Anim. Genet. 2019. [Google Scholar] [CrossRef]
  51. McPherron, A.C.; Lee, S.J. Double muscling in cattle due to mutations in the myostatin gene. Proc. Natl. Acad. Sci. USA 1997, 94, 12457–12461. [Google Scholar] [CrossRef] [Green Version]
  52. Miranda, M.E.; Aigues, M.Y.; Boscher, M.Y.; Menissier, F.; Cortes, O.; Nunner, S. Simultaneous genotyping to detect myostatin gene polymorphism in beef cattle breeds. J. Anim. Breed. Genet. 2002, 119, 361–366. [Google Scholar] [CrossRef] [Green Version]
  53. Trendelenburg, A.U.; Meyer, A.; Rohner, D.; Boyle, J.; Hatakeya, A.S.; Glass, D.J. Myostatin reduces Akt/TORC1/p70S6K signaling, inhibiting myoblast differentiation and myotube size. Am. J. Physiol. Cell Physiol. 2009, 296, 1258–1270. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  54. Hifzan, R.M.; Idris, I.; Yaakub, H. Growth pattern for body weight, height at withers and body length of kalahari red goats. Pak. J. Pharm. 2015, 18, 200–203. [Google Scholar] [CrossRef] [PubMed]
Animals 10 00168 g001 550
Figure 1. Agarose electrophoresis of MSTN 5 bp indel (A), and sequencing chromas for the 5 bp indel in the MSTN gene (B). Sequencing chromas showed homozygotic deletion type (DD) and heterozygote type (ID).
Figure 1. Agarose electrophoresis of MSTN 5 bp indel (A), and sequencing chromas for the 5 bp indel in the MSTN gene (B). Sequencing chromas showed homozygotic deletion type (DD) and heterozygote type (ID).
Animals 10 00168 g001
Animals 10 00168 g002 550
Figure 2. Association of the 5 bp indel within the MSTN gene with body height, height at hip cross, and chest width index in kids of SBWC goats.
Figure 2. Association of the 5 bp indel within the MSTN gene with body height, height at hip cross, and chest width index in kids of SBWC goats.
Animals 10 00168 g002
Table
Table 1. Genetic parameters for the 5 bp indel of the SBWC goat MSTN (Myostatin) gene.
Table 1. Genetic parameters for the 5 bp indel of the SBWC goat MSTN (Myostatin) gene.
Sample SizesFrequenciesHomoHeteroNePICHWE p-Value
GenotypesAlleles
N = 1074DD 0.890D 0.9430.8960.1041.1160.0980.055
ID 0.110I 0.057
II 0.000
Note: HWE: Hardy–Weinberg equilibrium; Homo: Homozygosity; Hetero: Heterozygosity; Ne: Effective allele numbers; PIC: Polymorphism information content; SBWC: Shaanbei White Cashmere goat.
Table
Table 2. Relationship between the 5 bp indel locus within the MSTN gene and the growth traits in kids of SBWC goats (LSM a ± SE) (p < 0.05).
Table 2. Relationship between the 5 bp indel locus within the MSTN gene and the growth traits in kids of SBWC goats (LSM a ± SE) (p < 0.05).
Growth TraitsObserved Genotypes (LSM a ± SE)p-Values
DDID
Body Height (cm)53.27 a ± 0.14 (n = 548)52.20 b ± 0.50 (n = 54)0.02
Height at Hip Cross (cm)55.45 a ± 0.14 (n = 548)54.54 b ± 0.48 (n = 54)0.05
Body Length (cm)61.70 ± 0.20 (n = 548)60.73 ± 0.48 (n = 54)0.13
Hip Width (cm)12.85 ± 0.07 (n = 548)12.96 ± 0.30 (n = 54)0.72
Chest Width (cm)18.48 ± 0.11 (n = 548)19.10 ± 0.36 (n = 54)0.09
Cannon bone Circumference (cm)7.38 ± 0.03 (n = 548)7.32 ± 0.10 (n = 54)0.47
Chest Depth (cm)25.44 ± 0.11 (n = 548)25.25 ± 0.31 (n = 54)0.54
Body Weight (kg)36.22 ± 0.93 (n = 73)35.70 ± 2.36 (n = 11)0.84
Heart Girth (cm)72.11 ± 0.31 (n = 548)72.30 ± 0.94 (n = 54)0.92
Body Trunk index118.36 ± 1.77 (n = 548)119.07 ± 1.29 (n = 54)0.90
Body Length index115.98 ± 0.47 (n = 548)116.89 ± 1.42 (n = 54)0.57
Heart Girth index135.67 ± 0.69 (n = 548)139.27 ± 2.41 (n = 54)0.12
Cannon bone Circumference index13.89 ± 0.07 (n = 548)14.08 ± 0.22 (n = 54)0.43
Chest Width index72.79 a ± 0.42 (n = 548)75.67 b ± 1.09 (n = 54)0.04
Hip Width index147.57 ± 2.97 (n = 548)149.50 ± 2.89 (n = 54)0.84
Note: Cells with different letters. a,b differed significantly (p < 0.05).

Share and Cite

MDPI and ACS Style

Bi, Y.; Feng, B.; Wang, Z.; Zhu, H.; Qu, L.; Lan, X.; Pan, C.; Song, X. Myostatin (MSTN) Gene Indel Variation and Its Associations with Body Traits in Shaanbei White Cashmere Goat. Animals 2020, 10, 168. https://doi.org/10.3390/ani10010168

AMA Style

Bi Y, Feng B, Wang Z, Zhu H, Qu L, Lan X, Pan C, Song X. Myostatin (MSTN) Gene Indel Variation and Its Associations with Body Traits in Shaanbei White Cashmere Goat. Animals. 2020; 10(1):168. https://doi.org/10.3390/ani10010168

Chicago/Turabian Style

Bi, Yi, Bo Feng, Zhen Wang, Haijing Zhu, Lei Qu, Xianyong Lan, Chuanying Pan, and Xiaoyue Song. 2020. "Myostatin (MSTN) Gene Indel Variation and Its Associations with Body Traits in Shaanbei White Cashmere Goat" Animals 10, no. 1: 168. https://doi.org/10.3390/ani10010168

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Metrics

Back to TopTop